Seasonal effects on behavioral traits of stabled Criollo stallions during semen collection

Abstract

semen filtration for their sperm retention efficiency and their effects on sperm quality parameters. Fifteen ejaculates from four stallions were collected and immediately extended with skim milk 1:1 (v/v). Extended semen was adjusted to a final concentration up to 50 x 106 sptz/mL using a Neubauer chamber. Three aliquots of each ejaculate were submitted to the following procedures: (a Cent) centrifugation of 20 mL of extended semen in a 50 mL conical tube at 600 x g for 10 min, (b Cushion) centrifugation at 1000 x g for 20min of 20mL of extended semen in a 50 mL conical tube with 1 mL of cushion solution (Eqcellsire component B, IMV, France) at the bottom, and (c Filter) filtration with commercial filter (Sperm Filter , Botupharma, Brazil) with a hydrophilic synthetic membrane that by gravity allows the seminal plasma to pass through and not the sperm, due to pore diameter. Following centrifugation supernate was aspirated and, when present, the cushion fluid was removed. Sperm pellets and filtered semenwas resuspended with skimmilk at 25 x 106 sperm/mL and immediately evaluated for total motility, progressive motility, velocity, membrane funcionality (HOST) and membrane integrity (CFDA/PI). Data were analyzed by one-way ANOVA andmeans compared by the Tukey test at 5% level of significance. The sperm recovery rate of each method was calculated dividing the total number of sperm recovered by the initial total sperm number of each aliquot. The results are shown in Table 1.